It is considered that reactive oxygen species (ROS) derived from immunocytes such as neutrophils and phagocytes not only biophylactically acts on the invaded pathogen (Babior, B. M., N. Engl. J. Med., 298, 659-668, 721-725, 1978), but also destructively on tissue in inflammation or circulation disorders (Weiss, S. J., N. Engl. J. Med., 320, 365-376, 1989). The main source of ROS production by neutrophils is NAD(P)H oxidase (Hallett, M. B. et al., Immunology Today, 16, 264-2681 1995), and it has been indicated that inhibition of neutrophil NAD(P)H oxidase can decrease organopathy in diseases involving neutrophils, such as inflammatory disease and circulation disorders (Schmid-Schonbein, G. W. et al., Physiology and pathology of leukocyte adherence, New York, Oxford University Press, 1995).
On the other hand, it has been previously reported that non-phagocytes such as smooth muscle cells, fibroblasts and vascular endothelial cells also can produce superoxide anion (O2−) dependant on NADPH or NADH and the potential association with cell functions, such as cell proliferation, hyperpermeability, and contraction and relaxation has been suggested (Griendling, K. K. et al., Circ.Res., 86, 494-501, 2000). Oxygen itself was first considered to be almost the same as neutrophil NAD(P)H oxidase. In recent years, the genes of the isozyme of gp91-phox, as a membrane-constituent factor of neutrophil NAD(P)H oxidase, was successively cloned. At present, Duox (dual oxidase) is known as an isozyme having five kinds of Nox from Nox 1 to Nox 5, peroxidase activity and clearly forms the Nox-Duox family, suggesting its potential involvement in various tissue and cell functions and the onset of diseases (Lambeth, J. D., Curr. Opin. Hematol., 9, 11-17, 2002).
The NAD(P)H oxidase of vascular smooth muscle cells and vascular endothelial cells is activated by many stimulations such as blood pressure regulatory hormones including angiotensin II (Ang II), cytokine, thrombin, PDGF, insulin, mechanical stimulation, hyperglycemia and hyperlipemia etc., predicting its involvement in various cardiovascular diseases. The spontaneous hypertensive rat model or the hypertensive rat model by the continuous administration of Ang II, has been reported to result in an increase in O2− production in blood vessel walls via NAD(P)H oxidase and the suppression of the blood pressure increase by the inhibition of NAD(P)H oxidase (Chen, X. et al., Hypertension, 38, 606-611, 2001; Rey, F. E. et al., Circ. Res., 89., 408-414, 2001), suggesting the potential involvement of NAD(P)H oxidase in blood pressure regulation.
Arteriosclerosis lesions are a chronic inflammatory multiplicative change in blood vessels and ROS produced in blood vessel walls plays an important role in its onset and development. In the p47phox knockout mouse, which fails to express a cytoplasmic component of NAD(P)H oxidase, the development of arteriosclerosis lesions has been reported to be inhibited when the mice are subjected to a high cholesterol load (Stokes, K. Y. et al. Circ. Res., 88, 499-505, 2001; Barry-Lane, P. A. et al., J. Clin. Invest., 108, 1513-1522, 2001). ROS is involved in the neointima growth after balloon injury and induces vascular restenosis. In recent years, the increase in NAD(P)H oxidase activity in vascular wall after balloon injury was reported (Shi, Y. et al., Arterioscler. Thromb. Vasc. Biol., 21, 739-745, 2001; Szocs, K. et al., Arterioscler. Thromb. Vasc. Biol., 22, 21-27, 2002). It was also reported that the NAD(P)H oxidase hypoactivity by C242T genetic variation of p22phox, as a cell membrane component, correlated with a decrease in the incidence rate of coronary artery disease (Inoue, N. et al., Circulation, 97, 135-137, 1998; Cai, H. et al., Eur. J. Clin. Invest., 29, 744-748, 1999; Cahilly, C. et al., Circ. Res., 86, 391-395, 2000). These reports indicate the potential involvement of NAD(P)H oxidase in the onset and development of arteriosclerosis and coronary artery diseases.
The potential involvement of ROS in the onset and development of diabetic complications has been indicated. It has been reported that the stimulation by hyperglycemia or glycated protein enhances oxidative stress via NAD(P)H oxidase in vascular endothelial cells, smooth muscle cells, etc. (Inoguchi, T. et al. Diabetes, 49, 1939-1945, 2000; Hink, U. et al. Circ. Res., 88, E14-E22, 2001; Wautier, M. et al., Am. J. Physiol., 280, E685-E694, 2001). The correlation between the increase in NAD(P)H oxidase activity and the injury of retinal vascular endothelial cells was also reported in retinal vessels of a diabetic model rat (Ellis, E. A. et al., Free Radic. Biol. Med., 24, 111-120, 1998).
For cerebral circulation disorders like stroke, it has been reported that leukocytes are involved in tissue injury (Hartl, R. et al., J. Cereb. Blood Flow Metab., 16, 1108-1119, 1996). The reduction of cerebral ischemia lesions has been reported in mice defective in neutrophil NAD(P)H oxidase activity (Walder, C. E. et al., Stroke, 28, 2252-2258, 1997). It also has been reported that stimulation by ischemia, inflammation, β-Amyloid, etc. could induce neuronotoxicity by activating NAD(P)H oxidase of microglia cells (Spranger, M. et al. J. Cereb. Blood Flow Metab., 18, 674-678, 1998; Vianca, V. D. et al., J. Biol. Chem., 274, 15493-15499, 1999; Green, S. P. et al., J. Cereb. Blood Flow Metab., 21, 374-384, 2001). These results suggest the potential involvement of NAD(P)H oxidase in stroke and neurodegenerative diseases.
ROS produced from NAD(P)H oxidase is involved in cell proliferation and vascularization, suggesting an association with tumor hyperplasia (Arnold, R. S. et al., Proc. Natl. Acad. Sci. USA, 98, 5550-5555, 2001; Arbiser, J. L. et al., Proc. Natl. Acad. Sci. USA, 99, 715-720, 2002).
Besides the aforesaid, NAD(P)H oxidase activity has been reported in the kidney cells, gastric cells, fat cells, chondrocytes, etc., raising the association with cell function. As stated above, NAD(P)H oxidase is widely associated with the onset and development of the diseases based on inflammation, circulation disorders and enhancement of proliferation activity, etc., i.e. hypertension, diabetic complications, arteriosclerosis, coronary artery disease, stroke, ischemic disease, neurodegenerative diseases, pulmonary circulation disorders, nephritis, arthritis, inflammatory diseases, cancer, etc. There is a possibility that NAD(P)H oxidase inhibitors can restrain these diseases.
The following compounds having a pyrazolo-[1,5-a]-pyrimidine backbone are known in the art.
Japanese Laid-open Patent Publication No. 5-112571 discloses the following compound:
wherein, R1 is hydrogen, or OH;                R2 is hydrogen, lower alkoxycarbonyl, lower alkoxy, halogen, lower alkyl, —CONHR6 (R6 is hydrogen, phenyl that may have a halogen atom, lower alkyl), or the like;        R3 is hydrogen, OH, lower alkyl, or the like;        
R5 is hydrogen, lower alkyl, lower alkoxy lower alkyl, or halogenated lower alkyl; and                R4 is hydrogen, lower alkyl, or lower alkoxy. This publication discloses that this compound inhibits the expression of androgen functions, and may be used for the treatment of enlarged prostate, female hirsutism, male baldness, acne, and the like.        
WO 00/59908 discloses the following compound:
wherein, R3 is (substituted) aryl, or (substituted) heteroaryl; and                R4 and R5 are hydrogen, halogen, CN, alkyl, alkenyl, alkynyl, cycloalkyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, amino, alkylamino, or (substituted) phenyl. This compound has corticotropin releasing factor receptor antagonizing functions, and its uses include mental diseases, nervous diseases, anxiety, trauma stress, eating disorders, circulatory diseases, and the like.        
Japanese Laid-open Patent Publication No. 10-101672 discloses the following compound:
wherein, R1 is hydrogen, (substituted) lower alkyl, cycloalkyl, thienyl, furyl, lower alkenyl, or (substituted) phenyl; and                R5 is hydrogen, or lower alkyl. This compound is used as an adenosine enhancer. Its uses includes the treatment of cardiac infarction, and brain infarction.        
Japanese Laid-open Patent Publication No. 7-157485 discloses the following compound:
wherein, R1 is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, lower alkoxy, or lower alkylthio; and                X, Y, Z are N or CR3. This compound is an angiotensin II antagonist. Its uses include the treatment of circulatory diseases such as stroke.        
EP 0328700A1 discloses the following compound:
The uses of this compound include the treatment of cerebral circulatory diseases.
WO 00/53605 discloses the following compound:
wherein, X is CH or N;                R1 and R3 are hydrogen, alkyl, alkenyl, alkynyl, aryl, halo, OH, or heterocyclyl; and        R5 is hydrogen, alkyl, OH, O-alkyl, halo, amino, or nitro. This compound has tyrosine kinase inhibitory action. Its uses include the treatment of cancers, vascularization, diabetic complications, inflammation, and the like.        
WO98/54093 discloses the following compound:
wherein, R1 is hydrogen, (substituted) alkyl, cycloalkyl, aryl, (substituted) heterocyclyl, halo, OH, or (substituted) heteroaryl;                R2 and R3 are hydrogen, alkyl, aryl, cycloalkyl, OH, halo, amino, or nitro;        R4 is hydrogen, (substituted) alkyl, cycloalkyl, alkoxy, (substituted) alkenyl, (substituted) alkynyl, (substituted) aryl, (substituted) heterocyclyl, alkoxy-NRR, NO2, OH, NH2, or (substituted) heteroaryl; and        R5 is hydrogen, alkyl, alkoxy, OH, halo, NO2, or NH2.This compound has tyrosine kinase inhibitory action. Its uses include the treatment of cancers, vascularization, diabetic complications, inflammation, and the like.        
Japanese Laid-open Patent Publication No. 4-270285 discloses the following compound:
wherein, Y is a lower alkylene, or lower alkenylene; and                Z is substituted acetyl, heterocyclic, or the like. This compound inhibits HMGCOA reducing enzyme. Its uses include the treatment of hyperlipemia.        
WO00/44754 discloses the following compound:
wherein, R2 and R3 are hydrogen, halogen, (substituted) alkyl, (substituted) alkenyl, (substituted) aryl, (substituted) aralkyl, or (substituted) heterocyclic group, or together form an alkylene group.                X is N or CR4. This compound inhibits fat accumulation. Its uses include the treatment of obesity, diabetes, and hypertension.        
Japanese Patent Publication No. 2000-38350 discloses the following compound:
wherein, E is N, or CR9 (R9 is hydrogen, alkyl, halogen, cyano, hydroxy, or alkoxy);                R1 is hydrogen, alkyl, cycloalkyl, alkoxy, (alkyl)amino, aryl, or heteroaryl;        J is NR2R3, or OR10; and        G is C or N. The heterocycles of the A ring include        
This compound has corticotropin releasing factor (CRF) receptor antagonizing functions. Its uses include the treatment of diabetes.
Japanese Laid-open Patent Publication No. 9-169762 discloses the following compound:
wherein, R5 is carboxy, or lower alkoxycarboxy, (substituted) carbamoyl (the substituent is lower alkyl, or phenyl lower alkyl); and                n is 1-5. The functions of this compound are unknown. Its uses include pain relief, lowering blood sugar level, and the treatment of inflammation, bacterial infection, cancers, and the like.        
Khim.-Farm. Zh (1995), 29 (4), 37-38 discloses (2,5-dimethyl pyrazolo-[1,5-a]-pyrimidine-7-yl) succinic acid. Its uses include the treatment of diabetes.
The purpose of the present invention is to provide novel compounds that inhibit NAD(P)H oxydase, and compositions comprising the compound. Another purpose of the present invention is to provide pharmaceutical compositions (including quasi-drugs), animal drug (livestock drugs, veterinary drugs, fishery drugs, and the like) compositions, as well as diagnostic drugs that are used to diagnose NAD(P)H-related diseases.
A further purpose of the present invention is to treat or prevent diseases due to inflammation, circulatory disorders, enhanced proliferation activities, and the like, i.e., hypertension, diabetic complications, arteriosclerosis, coronary artery disorders, strokes, ischemic heart disease, neurodegenerative diseases, pulmonary circulation disorders, nephritis, arthritis, inflammatory diseases, cancers and the like.